The granzymes are highly conserved group of serine proteases, with five members (A, B, H, K and M) in humans and ten members (A-G, K, M-N) in mice (Sattar R. et al. Biochem Biophys Res Commun 308, 726-35 (2003). Granzyme B (GrB or cytotoxic T-lymphocyte (CTL)-associated gene transcript-1—Brunet J. F. et al. Nature 322, 268-71 (1986)), has been reported as being involved in anti-viral and anti-tumour functions, and is associated with autoimmunity, transplant rejection, graft-versus-host disease, and thymocyte development (Barry M. & Bleackley R C. Nat Rev Immunol 2, 401-9 (2002)). Granzyme A (GrA) is also involved it immune-mediated killing, and is expressed by both innate and adaptive immune cytotoxic cells.
GrB is reported to have a contribution to CTL-mediated target cell apoptosis. GrB-deficient mice possess a normal phenotype, with the exception of a slightly reduced CTL-mediated target cell apoptosis, anti-viral responses and tumour cell clearance (Revell P A. et al. J Immunol 174, 2124-31 (2005); and Heusel J W. et al. Cell 76, 977-87 (1994)), suggesting a redundancy in immune mediated cells removal. GrB-deficient recipient mice exhibit reduced allograft vasculopathy (Choy J. C. et al. Am J Transplant 5, 494-9 (2005)), and its deficiency in mice leads to increased susceptibility to allergen-induced asthma (Devadas, S. et al. Immunity 25, 237-47 (2006)).
Buzza M. S. et al. report that plasma GrA levels in normal individuals are between 15-35 pg/ml and plasma GrB levels in normal individuals are up to 15 pg/ml (Biol. Chem. 387:827-837 (2006)). Skjelland, et al. teach that plasma levels of granzyme B are increased in patients with lipid rich carotid plaques, but also teach that normal plasma levels of GrB can be up to 100 pg/ml, while patterns with unstable plaques have plasma levels of GrB over about 100 pg/ml (and up to about 650 pg/ml) and patients with stable plaques have plasma levels of GrB between about 25 pg/ml and about 400 pg/ml (Atherosclerosis, 195:e142-e146 (2007)). Furthermore, GrB levels have been measured in the supernatant of cultured peripheral blood mononuclear cells isolated from patients at about 40 pg/ml for patients with unstable angina pectoris and at about 18 pg/ml for patients with stable angina pectoris (Tsuru R. et al. Heart 94:305-310 (2008) e-published Jun. 25, 2007). GrB has also been measured in rheumatoid arthritis patients (Goldbach-Mansky et al. Ann Rheum Dis. 64:715-721 (2005); Kraan et al. Ann Rheum Dis 63:483-488 (2004); Villanueva et al. Arthritis Res Ther 7:R30-R37 (2005)). Choy J C. et al. reported increased levels of GrB in patients with advanced atherosclerosis (Mod Pathol 16, 460-70 (2003)). GrB has also been associated with aortic aneurisms and with atherosclerosis plaque destabilization (Choy et al. Arterioscler. Thromb. Vasc. Biol.; 24; 2245-2250, (2004)). Kim et al., show that macrophages express granzyme B in the lesion areas of atherosclerosis and rheumatoid arthritis (Immunology Letters, 111, 57-65, (2007)). Increased GrB levels in Chronic Obstructive Pulmonary Disease (COPD) patients were reported in bronchoalveolar lavage (BAL) derived T-cells (Hodge et al. J. of COPD 3:179-187 (2006)). Also GrB produced protein fragments are reported in Sjögren's Syndrome patients (Huang et al. Clin Exp Immun 142:148-154 (2005)). Additionally, GrA and GrB are reported in BAL fluids from patients with inflammatory lung disease (Tremblay et al. J Immunology 165:3966-3969 (2000)). Thewissen et al. compares GrA and GrB levels in rheumatoid arthritis (RA), multiple sclerosis (MS), and between healthy individuals, and reports no change in GrA levels between healthy patients and RA or MS patients for GrA and reports a decrease in GrB levels for MS patients relative to healthy patients (Clinical Immunology 123:209-218 (2007)). GrA released in the brain may be associated with autoimmune disorders of the nervous system (Suidan et al. PNAS 91:8112-8116 (1994)). Vernooy et al. report increased GrA expression in type II pneumocytes of patients with severe COPD (Am J Respir Crit Care Med 175:464-472 (2007)). Immunodiagnostic methods for Granzymes A and B are also known (for example WO 99/54737).